SCHEME FOR THE IDENTIFICATION OF UNKOWN SUBSTANCES

Physical properties Chemical properties

Presented by Dr Nada Soliman

Number of the unknown

q

name number...

Physical properties:

1. Shape 2. Color 3. Odor 4. Taste 5. Solubility 6. Reaction 7. Heat dry test

q

Chemical properties test observation result

unknown possibilities: CHO

Reducing heat-coag non reducing heat-coagulable

protein
non

Glucose peptone Maltose Lactose (considered As one solution)

starch gelatin

sucrose

albumin and globuline

Physical Properties
1.

Shape
A. B.

Powder e.g. starch. Crystals e.g. sugars. Color. Odor. Taste: The unknown substance may be:

2.

3.

4.

A. B.

Sweet e.g. sugars Tasteless e.g. starch

5.

Solubility:
A. B.

Cold water: e.g. sugars urea,.etc. Hot water: e.g. starch and gelatin

6. 7.

Reaction Heat dry test: Heat a small portion of the unknown substance in a dry test tube
A. B. C.

Odor of burnt bread e.g. starch. Odor of caramel e.g. sugars. Odor of burnt hair e.g. Protein

Chemical Properties
We have 3 solutions of protein, one of them : egg white solution (source of albumine and globuline)is a natural solution which contain beside protein, CHO So we start the scheme with biuret test (test specific for protein)

Biuret test

Heat coagulation test

Millons test

ick to edit Master text styles Second level Third level Fourth level Fifth level

test
1. 2.

Observation
Violet color White ppt in the upper part of the test tube ppt with saturated (NH4)2SO4 solution ppt with fully saturated (NH4)2SO4 solution Brick red ppt

Result Protein solution heat coagulable protein (albumin & globulin) Globulin Albumin Protein contain Tyrosine

Biuret test Heat coagulation test Salting out

1.

1.

Millons test

Biuret test

Heat coagulation test

Millons test

test
1. 2.

Observation

result

Biuret test Heat Coagulation test Millons test

3.

Violet color No white ppt in the upper part of the test tube Brick red ppt Black ppt

3.

Sulfur test

Protein solution non-heat coagulable protein (peptone or gelatin) Protein containing tyrosine: peptone Proteine containing sulfur containing aa (cystine et cysteine): peptone

Biuret test

Heat coagulation test

Millons test

Test
1. 2.

Observation

Result

Biuret test Heat coagulation test

Violet color No white ppt in the upper part of the test tube No brick red ppt No Black ppt

Protein solution non-heat coagulable protein (peptone or gelatin) Protein doesnt contain tyrosine: gelatin Protein doesnt contain cystine or cysteine: gelatin

1.

Millons test

1.

Sulfur test

biuret test violet color no violet color (protein) (not protein) heat coagulation test whit ppt no white ppt (heat- coagulable

Biuret test

molisch test

Iodine test

test
1. 2. 3. 4.

observation

result

Biuret test Molisch test Iodine test Fehling after hydrolysis

No violet color Reddish violet ring Deep blue Color Red ppt

Not protein CHO solution starch Products of hydrolysis of starch have free anomeric carbon atoms

Biuret test

Molisch test

Iodine test

Fehling test

Essai
1. 2. 3. 4.

Observation

Resultat

Biuret test Molisch test Iodine test Fehling test Osazone test

No violet color Reddish violet ring No Deep blue Color Red ppt Needle- shaped crystals Plate- shaped crystals Rosette- shaped crystals

1.

Not protein CHO solution Not starch Reducing sugar ( glucose, maltose ou lactose) Glucose Maltose lactose

Biuret test

Molisch test

Iodine test

Fehling test

Seliwanoff test

Essai
1. 2. 3. 4.

Observation

Result

Biuret test Molisch test Iodine test Fehling test

No violet color Reddish violet ring No Deep blue Color No Red ppt Red onion peel color Red ppt

1. Seliwanoff test 1. Fehling after hydrolysis

Not protein CHO solution Not starch Non Reducing sugar (sucrose) Sucrose Products of hydrolysis of sucrose have free anomeric carbon atoms

molisch test reddish

violet ring no reddish violet ring (CHO) (not CHO)